In Vitro Transcription Systems from BY-2 Cells

Abstract

The in vitro transcription system is a powerful method to dissect molecular mechanisms of transcription of genes in question, usually performed in cellular or nuclear extracts containing transcription apparatuses with exogenous DNA templates and four nucleoside triphosphate (NTP) substrates. The first eukaryotic system was reported by Weil et al. (1979). They carried out in vitro transcription using a cytosolic extract and purified Pol II from HeLa cells. Manley et al. (1980) followed with in vitro assays using a whole cell extract from HeLa cells. In 1983, Dignam et al. (1983) established a standard procedure to prepare nuclear extracts from HeLa cells. Based on these efforts, many other in vitro systems have been established from different eukaryotic species, for example, yeast, mouse, and fruit fly. Since then, many significant discoveries have been made with these in vitro transcription systems.